Blastocyst Injection of Wild Type Embryonic Stem Cells Induces Global Corrections in Mdx Mice

Stillwell, Elizabeth E.; Vitale, Joseph M.; Zhao, Qingshi; Beck, Amanda J.; Schneider, J; Khadim, Farah; Elson, Genie; Altaf, Aneela; Yehia, Ghassan; Dong, Jiahui; Liu, Jing; Mark, Willie; Bhaumik, Mantu; Grange, Robert W.; Fraidenraich, Diego

doi:10.1371/journal.pone.0004759

Cited by 31 publications

(74 citation statements)

References 41 publications

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“…In our previous models with mdx mice, there was a noticeable lack of uniformity of dystrophin-positive fibers within the rescued muscle (Stillwell et al, 2009;Beck et al, 2011). However, in WT/SGd chimeras, dystrophin was detected in all fibers, but its overall staining intensity appeared to vary (Fig.…”

Section: Resultsmentioning

confidence: 71%

“…However, in previous reports, we showed that it takes approximately10-30% ESC (or iPSC) incorporation to rescue mdx mice, which are predisposed to DMD (Stillwell et al, 2009;Beck et al, 2011). This disparity could be because of several factors linked to differences between the mdx and SGd-KO MD mouse models and/or with their respective interactions with the ESCs.…”

Section: Discussionmentioning

confidence: 77%

“…At 3 weeks of age, the degree of ESC incorporation, or chimerism, was estimated by performing X-gal staining of sections of tail biopsies and semiquantitative (semi-Q) genomic PCR for both the WT and mutant SGd genes, as described previously (Stillwell et al, 2009;Beck et al, 2011). We analyzed four mice with ,5% ESC incorporation, one mouse with 10% incorporation, two with 40-60% incorporation and two with 60-80% incorporation.…”

Section: Resultsmentioning

confidence: 99%

“…S1), and by immunofluorescence analysis of the heart for the SGC (supplementary material Fig. S2Q-T) (Stillwell et al, 2009;Beck et al, 2011).…”

Section: Resultsmentioning

confidence: 99%

“…Both forms of PSCs were incorporated at low percentages into the resultant chimeras and supplied dystrophin to rescue the mdx but not the mdx:utrophin KO muscle tissue, although in both chimeric models there was a recovery of fat mass. This suggests that other tissues also play a role in the development of MD, and that the PSCs exert corrections globally (Stillwell et al, 2009;Beck et al, 2011). We wanted to study whether the ESC rescue of DMD accomplished in mdx mice with low chimerism also operated in other models of MD, such as LGMD-2F, which is recapitulated in SGd-KO mice.…”

Section: Introductionmentioning

confidence: 99%

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Dystrophin-compromised sarcoglycan-δ knockout diaphragm requires full wild type embryonic stem cell reconstitution for correction

Vitale

Schneider

Beck

et al. 2012

Journal of Cell Science

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SummaryLimb-girdle muscular dystrophy-2F (LGMD-2F) is an incurable degenerative muscle disorder caused by a mutation in the sarcoglycan-d (SGd)-encoding gene (SGCD in humans). The lack of SGd results in the complete disruption of the sarcoglycan complex (SGC) in the skeletal and cardiac muscle within the larger dystrophin-glycoprotein complex (DGC). The long-term consequences of SG ablation on other members of the DGC are currently unknown. We produced mosaic mice through the injection of wild-type (WT) embryonic stem cells (ESCs) into SGd-knockout (KO) blastocysts. ESC-derived SGd was supplied to the sarcolemma of 18-month-old chimeric muscle, which resulted in the restoration of the SGC. Despite SGC rescue, and contrary to previous observations obtained with WT/mdx chimeras (a mouse rescue paradigm for Duchenne muscular dystrophy), low levels of ESC incorporation were insufficient to produce histological corrections in SGd-KO skeletal muscle or heart. The inefficient process of ESC rescue was more evident in the SGd-KO diaphragm, which had reduced levels of dystrophin and no compensatory utrophin, and needed almost full WT ESC reconstitution for histological improvement. The results suggest that the SGd-KO mouse model of LGMD is not amenable to ESC treatment.

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Section: Resultsmentioning

confidence: 71%

Section: Discussionmentioning

confidence: 77%

Section: Resultsmentioning

confidence: 99%

“…S1), and by immunofluorescence analysis of the heart for the SGC (supplementary material Fig. S2Q-T) (Stillwell et al, 2009;Beck et al, 2011).…”

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Dystrophin-compromised sarcoglycan-δ knockout diaphragm requires full wild type embryonic stem cell reconstitution for correction

Vitale

Schneider

Beck

et al. 2012

Journal of Cell Science

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Four decades of leading‐edge research in the reproductive and developmental sciences: The Infant Primate Research Laboratory at the University of Washington National Primate Research Center

Burbacher

Grant

Worlein

et al. 2013

American J Primatol

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The Infant Primate Research Laboratory (IPRL) was established in the 1970s at the University of Washington as a visionary project of Dr. Gene (Jim) P. Sackett. Supported by a collaboration between the Washington National Primate Research Center and the Center on Human Health and Disability, the IPRL operates under the principle that learning more about the causes of abnormal development in macaque monkeys will provide important insights into mechanisms underlying childhood neurodevelopmental disorders. Over the past forty years, a broad range of research projects have been conducted at the IPRL. Some have described the normal expression of species-typical behaviors in nursery-reared macaques while others have focused on specific issues in perinatal medicine and research. This article will review the unique history of the IPRL and the scientific contributions produced by research conducted in the laboratory. Past and present investigations at the IPRL have explored the consequences of adverse early rearing, low-birth-weight, prematurity, epilepsy, chemical/drug exposure, viral infection, diarrheal disease, vaccine safety, assisted reproductive technologies and perinatal hypoxia on growth and development. New directions of investigation include the production of a transgenic primate model using our embryonic stem cell-based technology to better understand and treat heritable forms of human mental retardation such as fragile X.

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Generation of viable PDX1 gene‐edited founder pigs as providers of nonmosaics

Tanihara

Hirata

Nguyen

et al. 2020

Molecular Reproduction Devel

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Pancreatic duodenal homeobox 1 (PDX1) is a crucial gene for pancreas development during the fetal period. PDX1-modified pigs have the potential to be used as a model of diabetes mellitus. However, the severe health problems caused by the PDX1 mutation limit phenotypic studies of PDX1-modified pigs as diabetes models. In this study, we generated PDX1-modified pigs by the CRISPR/Cas9 system introduced into zygotes via electroporation and investigated the mosaicism, phenotypes, and inheritance of the resulting pigs. After the embryo transfer of PDX1-modified zygotes, nine mutant piglets were delivered. Two piglets were apancreatic biallelic mutants. For the other seven piglets, the ratio of mutant alleles to total alleles was 17.5-79.7%. Two mutant piglets with high mutation rates (67.7% and 79.7%) exhibited hypoplasia of the pancreas, whereas the other five piglets were healthy.One of the male mutant piglets was further analyzed. The ejaculated semen from the pig contained PDX1-mutant spermatozoa and the pig showed normal reproductive ability. In conclusion, the frequency of the PDX1 mutation is presumed to relate to pancreas formation, and PDX1 mutant founder pigs generated from zygotes introduced to the CRISPR/Cas9 system can serve as providers of nonmosaics to contribute to medical research on diabetes mellitus. K E Y W O R D SCRISPR/Cas9, diabetes mellitus, electroporation, IVF, PDX1-mutant pigs SUPPORTING INFORMATIONAdditional supporting information may be found online in the Supporting Information section.

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Blastocyst Injection of Wild Type Embryonic Stem Cells Induces Global Corrections in Mdx Mice

Cited by 31 publications

References 41 publications

Dystrophin-compromised sarcoglycan-δ knockout diaphragm requires full wild type embryonic stem cell reconstitution for correction

Dystrophin-compromised sarcoglycan-δ knockout diaphragm requires full wild type embryonic stem cell reconstitution for correction

Four decades of leading‐edge research in the reproductive and developmental sciences: The Infant Primate Research Laboratory at the University of Washington National Primate Research Center

Generation of viable PDX1 gene‐edited founder pigs as providers of nonmosaics

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